USB CycleSeq™ Thermostable DNA Polymerase
®
Cycle sequencing—Incorporates ddNTPs and dNTPs at equal
efficiencies, ensuring banding patterns are uniform and easily
read
n SNP genotyping—Single base extension using nucleotide
analogs
n USB CycleSeq Thermostable DNA Polymerase can be substituted for Thermo Sequenase DNA polymerase.
n
USB CycleSeq is comparable to GE Thermo Sequenase
GE Thermo USB CycleSeq
Sequenase Thermostable
DNA
DNA
Polymerase Polymerase
GATC
GATC
USB CycleSeq Thermostable DNA Polymerase is a thermostable,
DNA polymerase that is exonuclease-free and incorporates
ddNTPs with a marked level of efficiency over standard thermostable DNA polymerases. This allows for more even and easy-to-read sequence band patterns, making sequence anomalies easier to identify.
CycleSeq DNA Polymerase, like Thermo Sequenase DNA
Polymerase, includes Thermoplasma acidophilum inorganic
pyrophosphatase (TAP). TAP is thermostable and converts pyrophosphate to orthophosphate, a useful feature during cycle
sequencing reactions where pyrophosphates are produced.
USB CycleSeq Thermostable DNA Polymerase is useful for
cycle sequencing (Sanger sequencing), as it produces sequence
data with uniform band intensities which allows for longer and
more accurate sequence reads. It is also useful for primer extension protocols during SNP genotyping.
Properties:
CycleSeq Thermostable DNA Polymerase is a thermostable DNA
polymerase that incorporates dideoxynucleotides (ddNTPs) as
readily as deoxynucleotides (dNTPs).
Source:
Recombinant
Purity:
Greater than 95% pure as determined by SDS-PAGE.
Storage buffer:
20 mM Tris-HCl, pH 8.5, 0.1 mM EDTA, 1 mM DTT, 100 mM KCl,
0.053 unit/μl Thermoplasma acidophilum inorganic pyrophosphatase, 50% glycerol, and stabilizers.
Unit definition:
One unit of enzyme is defined as the amount of enzyme required
to catalyze the incorporation of 10 nmol of dNTPs into acidinsoluble form in 30 minutes at 74ºC under standard assay
conditions.
USB CycleSeq Polymerase sequencing results are comparable to GE
Thermo Sequenase. Cycle sequencing using a fluorescently-labeled primer
and 300 ng pUC19 template DNA was performed using either Thermo
Sequenase DNA Polymerase or CycleSeq Thermostable DNA Polymerase.
Analysis of both sequences shows equivalent quality, read length, and band intensity.
Assay conditions:
The reaction mixture contains 25 mM TAPS, pH 9.3, 50 mM KCl,
2 mM MgCl2, 1 mM b-ME, 200 μM dATP, 200 μM dGTP, 200 μM
dTTP, 100 μM dCTP, 0.05 µl [a-33P] dCTP (10 µCi/µl), 400 μg/ml
activated DNA, and CycleSeq Thermostable DNA Polymerase.
After incubation at 74°C for 10 minutes, acid-insoluble material
is determined (50 µl reaction volume).
Functional test:
Functionally tested to meet or exceed the specifications for
use in dye primer sequencing. Release specifications are based
on sequence length, band intensity, and sequence quality. The
sequence must also be free of background bands strong enough
to interfere with sequence interpretation.
Components:
CycleSeq Thermostable DNA Polymerase 1,000 units
1 ml CycleSeq Dilution Buffer
1 ml 10X CycleSeq Reaction Buffer
CycleSeq Thermostable DNA Polymerase 10,000 units
4 x 1 ml CycleSeq Dilution Buffer
2 x 1 ml 10X CycleSeq Reaction Buffer
Storage:
Shipped on dry ice. Store at -20°C.
CycleSeq Thermostable DNA Polymerase
Product code
79200
Concentration:
32 units/µl
Pack size
1,000 units
10,000 units
For Research Use Only. Not for use in diagnostic procedures.
Affymetrix and USB are registered trademarks of Affymetrix, Inc. CycleSeq is a trademark of Affymetrix, Inc. All other trademarks are the property of their respective owners.
Affymetrix, Inc.
USB® Products
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Fax: 800-535-0898 | 216-464-5075
[email protected]
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USB® Products
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Tel: +44 (0)1628 55 2600
Fax: +44 (0)1628 55 2675
[email protected]
© 2016 Affymetrix, Inc. All rights reserved.
P/N USB05291 Rev. 3
Affymetrix Pte Ltd.
USB® Products
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Keppel Logistics Building
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Tel: +65 63957310
Fax: +65 63957300
[email protected]
Rev 01/07/16